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Cd56 ecd

Manufactured by Beckman Coulter
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The CD56-ECD is a fluorochrome-conjugated antibody that binds to the CD56 antigen, which is expressed on natural killer (NK) cells, a subset of T cells, and some neural cells. The CD56-ECD antibody is used in flow cytometry applications for the identification and enumeration of CD56-positive cells.

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Lab products found in correlation

Cd3 pe cy7, by Thermo Fisher Scientific (2 mentions) Cd3 pe cy5, by Beckman Coulter (2 mentions) Cd45 fitc, by BD (1 mentions) Cd19 apc cy7, by BioLegend (1 mentions) Fcr blocking reagent, by Miltenyi Biotec (1 mentions) Lsr 2, by BD (1 mentions) Live dead cell viability stain, by Thermo Fisher Scientific (1 mentions) Cd146 fitc, by BioLegend (1 mentions) Cytokeratin fitc, by Thermo Fisher Scientific (1 mentions) Cd16 apc, by Thermo Fisher Scientific (1 mentions) Cd68 fitc, by Thermo Fisher Scientific (1 mentions) Cd14 v500, by BD (1 mentions) Tnf α fitc, by BD (1 mentions) Cd16 apc cy7, by BD (1 mentions) Ionomycin, by Merck Group (1 mentions) Live dead stain, by Thermo Fisher Scientific (1 mentions) Monensin, by Merck Group (1 mentions) Live dead stain, by BD (1 mentions) Rhil 12, by Miltenyi Biotec (1 mentions) Rhil 18, by R&D Systems (1 mentions)

Close spelling variants of this product

CYTO-STAT tetraCHROME CD45-FITC/CD56-RD1/CD19-ECD/CD3-PC5 (5 citations)

7 protocols using cd56 ecd

1

Quantifying TRAIL Receptor Expression

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Freshly isolated hHSC were stained with mAbs against CD19-APC-Cy7, CD146-FITC (BioLegend, London, UK), CD14-v500, CD45-FITC (BD Biosystems, Oxford, UK), CD68-FITC, Cytokeratin-FITC, CD3-PE-Cy7 (eBioscience, Hatfield, UK), CD56-ECD (Beckman Coulter, High Wycombe, UK), TRAIL-R1-PE, TRAIL-R2-PE, TRAIL-R3-PE and TRAIL-R4-PE (R&D Systems, Abingdon, UK) after FcR blocking reagent (Miltenyi Biotec, Surrey, UK) blocking. Appropriate isotype controls were used where necessary. NK cells were stained for CD56-ECD (Beckman Coulter, High Wycombe, UK), CD3-PE-Cy7 (eBioscience, Hatfield, UK), CD16-APC, TRAIL-PE. All cells were stained with Live/Dead® Cell viability stain (Invitrogen, California, USA). Data were acquired on BDTM LSR II flow cytometer (Beckton Dickinson, New Jersey, USA) and analysed using FlowJo (Treestar Oregon, USA).
Singh H.D., Otano I., Rombouts K., Singh K.P., Peppa D., Gill U.S., Böttcher K., Kennedy P.T., Oben J., Pinzani M., Walczak H., Fusai G., Rosenberg W.M, & Maini M.K. (2017). TRAIL regulatory receptors constrain human hepatic stellate cell apoptosis. Scientific Reports, 7, 5514.
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2

Cytokine Production in PBMCs

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For intracellular staining for IFN-γ production, PBMCs were incubated with 50 ng/ml of rhIL-12 (Miltenyi Biotec) and 50 ng/ml of rh-IL18 (R&D Systems) for 21 h at 37°C. One mM monensin (Sigma-Aldrich, Gillingham, UK) was added for the final 3 h. Cells were stained with antibodies to CD3-PE-Cy7 (eBioscience), CD16-APC-Cy7 (BD Biosciences) and CD56-ECD (Beckman Coulter) and subsequently fixed and permeabilized, followed by intracellular staining for IFN-γ-V450 (BD Biosciences). Dead cells were excluded by live/dead stain (Invitrogen).
For TNF-α production, PBMCs were stimulated with phorbol myristate acetate (PMA) (3 ng/ml) and ionomycin (100 ng/ml) (Sigma-Aldrich) for 3 h in the presence of 1 mM of monensin (Sigma-Aldrich). Cells were stained with antibodies to CD3-PE-Cy7 (eBioscience), CD16-APC-Cy7 (BD Biosciences) and CD56-ECD (Beckman Coulter) in the presence of live/dead stain followed by fixing, permeabilization and intracellular staining for TNF-α-FITC (BD Biosciences).
Heiberg I.L., Pallett L.J., Winther T.N., Høgh B., Maini M.K, & Peppa D. (2015). Defective natural killer cell anti-viral capacity in paediatric HBV infection. Clinical and Experimental Immunology, 179(3), 466-476.
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3

Comprehensive Immune Cell Profiling

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Killer-cell immunoglobulin-like receptor repertoire staining, functional assays, and data analysis were performed as described in detail elsewhere (3 (link)). In brief, freshly thawed PBMCs of patient and controls were stained with the following monoclonal antibodies: CD3-PE.Cy5, CD14-PE.Cy5, CD19-PE.Cy5, CD56-ECD, ILT2-PE, CD161-PE, CD7-PE-Cy7, and NKG2A-APC.AF750, all from Beckman Coulter; NKG2C-PE or biotin from R&D Systems; PLZF-PE, NKp30-APC, and NKp46-PE from Becton Dickinson; FcϵRγ1-FITC from Millipore; and CD57-PB from Biolegend. Dead cells were excluded by using the aqua live/dead kit (Invitrogen).
Béziat V., Sleiman M., Goodridge J.P., Kaarbø M., Liu L.L., Rollag H., Ljunggren H.G., Zimmer J, & Malmberg K.J. (2015). Polyclonal Expansion of NKG2C+ NK Cells in TAP-Deficient Patients. Frontiers in Immunology, 6, 507.
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4

Phenotypic Characterization of RSV-Infected NK Cells

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The following fluorochrome-conjugated monoclonal antibodies were used to phenotypically characterize (RSV-infected) NK cells: CD3-APCAF750 (UCHT1, Beckman Coulter), CD16-PacificOrange (3G8, Thermo Fisher), CD56-ECD (N901, Beckman Coulter), CD85j-PerCP-Cy5.5 (ILT2, LILRB1; GHI/75, BioLegend), CD161-APC (191B8, Miltenyi), CD158a-AF700 (KIR2DL1; 143211, R&D Systems), CD158a/h-PC5.5 (KIR2DL1/S1; EB6B, Beckman Coulter), CD158b1/b2,j-PC7 (KIR2DL2/L3/S2; GL183, Beckman Coulter), CD158e1-BV421 (KIR3DL1; DX9, BioLegend), CD159a-APC (NKG2A; Z199, Beckman Coulter), CD159c-PE (NKG2C; 134591, R&D Systems), CD244-AF700 (2B4; C1.7, BioLegend), CD314-APC (NKG2D; ON72, Beckman Coulter), CD335-PC7 (NKp46; BAB281, Beckman Coulter), CD336-PE (NKp44; Z231, Beckman Coulter), CD337-PerCP-Cy5.5 (NKp30; P30-15, BioLegend), RSV-G-FITC (131-2G, Millipore). Cells were measured using a Navios flow cytometer (Beckman Coulter).
van Erp E.A., Feyaerts D., Duijst M., Mulder H.L., Wicht O., Luytjes W., Ferwerda G, & van Kasteren P.B. (2018). Respiratory Syncytial Virus Infects Primary Neonatal and Adult Natural Killer Cells and Affects Their Antiviral Effector Function. The Journal of Infectious Diseases, 219(5), 723-733.
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5

Multicolor Flow Cytometry Panel

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The following monoclonal antibodies were used in the study: CD14-Horizon-V500, CD19-Horizon-V500 (BD Biosciences), CD3-PE-Cy5, CD56-ECD, CD57-PacificBlue, NKG2A-APC (Beckman Coulter), CD4-PE-Cy5, KIR3DL1-Alexa700 (Biolegend), KIR2DL1-biotin, KIR2DL3-FITC, NKG2C-PE (RnD systems), Aqua Dead Cell Stain Kit 405 nm, Strepavidin-Qdot-605, and Qdot-700 (Thermofisher).
Malone D.F., Lunemann S., Hengst J., Ljunggren H.G., Manns M.P., Sandberg J.K., Cornberg M., Wedemeyer H, & Björkström N.K. (2017). Cytomegalovirus-Driven Adaptive-Like Natural Killer Cell Expansions Are Unaffected by Concurrent Chronic Hepatitis Virus Infections. Frontiers in Immunology, 8, 525.
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6

Multiparametric Immunophenotyping of Immune Cells

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For the evaluation of surface antigen expression the following monoclonal antibodies (mAbs) were used: CD3-APC-A700, CD19-APC-A700, CD56-ECD, PC7 and APC-A700, CD11b-FITC, CD33-PC7, HLA-DR-PE, CD14-ECD, CD45-KrO, CD66b-APC, CD15-APC (all Beckman Coulter), CD107a-eFLUOR660 (Invitrogen), CD275-, CD155-, CD85j-, Ceacam1-, CD39-APC (Miltenyi biotec). For intracellular evaluation the following mAbs were used: anti-IFN-γ-PE (BD, biosciences), anti-TNF-α-eFluor450 (Invitrogen). For MDSC immunostaining a custom Duraclone platform (Beckman Coulter) was used in order to standardize the protocol. After staining procedures cells were acquired at Cytoflex S and LX (Beckman Coulter) and analyzed with Cytexpert software (v2.2, Beckman Coulter), and FlowJo 10 (Starlab).
Tumino N., Besi F., Martini S., Di Pace A.L., Munari E., Quatrini L., Pelosi A., Fiore P.F., Fiscon G., Paci P., Scordamaglia F., Covesnon M.G., Bogina G., Mingari M.C., Moretta L, & Vacca P. (2022). Polymorphonuclear Myeloid-Derived Suppressor Cells Are Abundant in Peripheral Blood of Cancer Patients and Suppress Natural Killer Cell Anti-Tumor Activity. Frontiers in Immunology, 12, 803014.
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7

Multiparametric Flow Cytometry Analysis

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All samples were treated with Fc receptor blocking reagent (Miltenyi Biotec) before staining. Surface staining was performed in 96 well plates (Sarstedt) in staining buffer of 50% PBS, 50% Brilliant Violet staining buffer (BD). Fixable live/dead stain (Life Technologies) was added to the staining buffer. Antibody staining was conducted for 15 min at 37°C in the dark before washing with PBS using the following monoclonal antibodies: CD3 PE-Cy7 and CD8 Alexa 700 (eBioscience), CD56 ECD (Beckman Coulter), NKG2D Alexa 488 (Biolegend). Cell viability was determined and dead cells excluded using fixable live/dead aqua stain (Invitrogen). Samples for were fixed in Cytofix (BD). Single fluorochrome compensation controls were made using compensation beads (BD). Compensation matrices were calculated initially in FACSDiva and edited where necessary in FlowJo X (TreeStar). Samples were acquired on LSR Fortessa (BD) and data analyzed in FlowJo X.
Easom N.J., Marks M., Jobe D., Gillmore R., Meyer T., Maini M.K, & Njie R. (2020). ULBP1 Is Elevated in Human Hepatocellular Carcinoma and Predicts Outcome. Frontiers in Oncology, 10, 971.
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